Molecular Biology

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    Protocols in Current Issue
    Cluster Analysis of Endogenous HER2 and HER3 Receptors in SKBR3 Cells
    The Human Epidermal Growth Factor Receptor (HER) family of receptor tyrosine kinases consists of four, single pass, transmembrane receptor homologs (HER1-4) that act to regulate many critical processes in normal and tumor cells. HER2 is ...
    Quantitative Transformation Efficiency Assay for Bacillus subtilis
    Authors:  Christian L. Loyo and Briana M. Burton, date: 12/05/2018, view: 163, Q&A: 0
    Bacillus subtilis (B. subtilis) is a model Gram-positive organism used to study a variety of physiological states and stress responses, one of which is the development of competence. Competence refers to the physiological state of ...
    Wheat Germ Agglutinin (WGA)-SDS-PAGE: A Novel Method for the Detection of O-GlcNAc-modified Proteins by Lectin Affinity Gel Electrophoresis
    Authors:  Ko Fujioka, Yuji Kubota and Mutsuhiro Takekawa, date: 12/05/2018, view: 129, Q&A: 0
    Diverse cytoplasmic and nuclear proteins dynamically change their molecular functions by O-linked β-N-acetylglucosamine (O-GlcNAc) modification on serine and/or threonine residues. Evaluation of the O-GlcNAcylation level of a specific protein, ...
    Cluster Analysis of Endogenous HER2 and HER3 Receptors in SKBR3 Cells
    [Abstract] The Human Epidermal Growth Factor Receptor (HER) family of receptor tyrosine kinases consists of four, single pass, transmembrane receptor homologs (HER1-4) that act to regulate many critical processes in normal and tumor cells. HER2 is overexpressed in many tumors, and the deregulated proliferation of cancerous cells is driven by cooperation with ...
    Quantitative Transformation Efficiency Assay for Bacillus subtilis
    Authors:  Christian L. Loyo and Briana M. Burton, date: 12/05/2018, view: 163, Q&A: 0
    [Abstract] Bacillus subtilis (B. subtilis) is a model Gram-positive organism used to study a variety of physiological states and stress responses, one of which is the development of competence. Competence refers to the physiological state of a cell which allows it to be transformed naturally. Through induction of competence, the efficiency ...
    Wheat Germ Agglutinin (WGA)-SDS-PAGE: A Novel Method for the Detection of O-GlcNAc-modified Proteins by Lectin Affinity Gel Electrophoresis
    Authors:  Ko Fujioka, Yuji Kubota and Mutsuhiro Takekawa, date: 12/05/2018, view: 129, Q&A: 0
    [Abstract] Diverse cytoplasmic and nuclear proteins dynamically change their molecular functions by O-linked β-N-acetylglucosamine (O-GlcNAc) modification on serine and/or threonine residues. Evaluation of the O-GlcNAcylation level of a specific protein, however, needs multiple and time-consuming steps if using conventional methods (e.g., ...
    Single-molecule Fluorescence in situ Hybridization (smFISH) for RNA Detection in Adherent Animal Cells
    Authors:  Gal Haimovich and Jeffrey E. Gerst, date: 11/05/2018, view: 582, Q&A: 1
    [Abstract] Transcription and RNA decay play critical roles in the process of gene expression and the ability to accurately measure cellular mRNA levels is essential for understanding this regulation. Here, we describe a single-molecule fluorescent in situ hybridization (smFISH) method (as performed in Haimovich et al., 2017) that detects ...
    mRNA Stability Assay Using Transcription Inhibition by Actinomycin D in Mouse Pluripotent Stem Cells
    Authors:  Madara Ratnadiwakara and Minna-Liisa Änkö, date: 11/05/2018, view: 243, Q&A: 0
    [Abstract] Gene expression is regulated through multiple steps at both transcriptional and post-transcriptional levels. The net abundance of mature mRNA species in cells is determined by the balance between transcription and degradation. Thus, the regulation of mRNA stability is a key post-transcriptional event that can greatly affect the net level of mRNAs ...
    RNA Immunoprecipitation Assay to Determine the Specificity of SRSF3 Binding to Nanog mRNA
    Authors:  Madara Ratnadiwakara and Minna-Liisa Änko, date: 11/05/2018, view: 223, Q&A: 0
    [Abstract] Mammalian cells express hundreds of RNA binding proteins (RBPs) that are essential regulators of RNA metabolism. RBP activity plays a central role in the control of gene expression programs and identification of RNA-protein interactions is critical for comprehensive understanding of gene regulation in cells. In recent years, various tools and ...
    Generation of Stable Expression Mammalian Cell Lines Using Lentivirus
    Authors:  Neha Tandon, Kaushik N. Thakkar, Edward L LaGory, Yu Liu and Amato J Giaccia, date: 11/05/2018, view: 281, Q&A: 0
    [Abstract] Lentiviruses are used very widely to generate stable expression mammalian cell lines. They are used for both gene down-regulation (by using shRNA) or for gene up-regulation (by using ORF of gene of interest). The technique of generating stable cell lines using 3rd generation lentivirus is very robust and it typically takes about 1-2 ...
    Protocol for in situ Proximity Ligation Assay (PLA) and Microscopy Analysis of Epidermal Growth Factor Receptor (EGFR) Homodimerization
    Authors:  Keiichi Ota and Taishi Harada, date: 11/05/2018, view: 341, Q&A: 0
    [Abstract] Oncogenic drivers play central roles in tumorigenesis as well as in tumor cell survival and proliferation. Mutations of the epidermal growth factor receptor gene (EGFR) that result in constitutive activation of the receptor tyrosine kinase have been identified as oncogenic drivers in a subset of non-small cell lung cancer (NSCLC). ...
    Kinetic Characterization of the Shigella Type Three Secretion System ATPase Spa47 Using α-32P ATP
    Authors:  Heather B. Case and Nicholas E. Dickenson, date: 11/05/2018, view: 204, Q&A: 0
    [Abstract] ATPases represent a diverse class of enzymes that utilize ATP hydrolysis to support critical biological functions such as driving ion pumps, providing mechanical work, unfolding/folding proteins, and supporting otherwise thermodynamically unfavorable chemical reactions. We have recently shown that the Shigella protein Spa47 is an ATPase ...
    Preparation and Purification of Proteins Secreted from Phytophthora sojae
    Authors:  Yeqiang Xia, Yan Wang and Yuanchao Wang, date: 10/20/2018, view: 520, Q&A: 0
    [Abstract] Phytophthora sojae, the causal agent of soybean root and stem rot, is responsible for enormous economic losses in soybean production. P. sojae secrets various effectors to reprogram host immunity. The plant apoplastic space is a major battleground in plant-pathogen interactions. Here we describe a protocol for purification and ...



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